• Title/Summary/Keyword: Activated T cell

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Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Induction of Apoptosis by Sageretia thea Branch Extracts through Activation of NF-κB Signaling Pathway in Human Colorectal Cancer Cells (상동나무(Sageretia thea) 가지추출물의 대장암세포에서 NF-κB 신호전달 활성화를 통한 세포사멸 유도활성)

  • Kim, Jeong Dong;Park, Su Bin;Eo, Hyun Ji;Park, Gwang Hun;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.33 no.5
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    • pp.428-435
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    • 2020
  • In this study, we evaluated the inhibitory effect against cell growth and potential molecular mechanism of 100% ethanol extracts of branch from Sageretia thea in human colorectal cancer cells, HCT116. Ethanol dose-dependently extracts of STB significantly suppressed the growth of HCT116 cells through apoptosis. STB activated NF-κB signaling pathway through IκB-α proteasomal degradation and inducing p65 accumulation in nucleus. The inhibition of GSK3β by LiCl didn't affect STB mediated degradation IκB-α but STB mediated p65 accumulation in nucleus. In addition, STB phosphorylated GSK3β. Based on these findings, STB may be a potential candidate for the development of anti-cancer agents for human colorectal cancer.

STUDY ON MUTATION OF RAS GENE IN DMBA INDUCED CARCINOMA OF HAMSTER BUCCAL POUCH (DMBA로 유도된 햄스터 협낭암종에서 ras 유전자 변이에 관한 연구)

  • Song, Sun-Chul;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.6
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    • pp.581-590
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    • 2000
  • Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Over 30 oncogenes can be activated by insertional mutagenesis, single point mutations, chromosomal translocations and gene amplification. The ras oncogenes have been detected in $15{\sim}20%$ of human tumors that include some of the most common forms of human neoplasia and are known to acquire their transforming properties by single point mutations in two domains of their coding sequences, most commonly in codons 12 and 61. The ras gene family consists of three functional genes, N-ras, K-ras and H-ras which encode highly similar proteins of 188 or 189 amino acid residues generically known as P21. ras proteins have been shown to bind GTP and GTP, and possess intrinsic GTPase activity. Experimental study was performed to observe the mutational change of the ras gene family and apply the results to the clinical activity. 36 Golden Syrian Hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek (control side) was treated with mineral oil as the same manner of the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were completely dissected by microdissection and DNA from both tissue were isolated by proteinase K/phenol/chloroform extraction. Segments of the K-ras and H-ras gene were amplified by PCR using the oligonucleotide primers corresponding to the homologous region (codon 12 and 61) of the hamster gene, and then confirmational change of ras genes was observed by SSCP and autosequencing analysis. The results were as follows : 1. Malignant lesion could be found in the experimental side from the experimental six weeks. 2. One hamster among six showed point mutation of the H-ras codon 12($G{\rightarrow}A$ transition) at the experimental 10 and 14 weeks. 3. One of six at 6 weeks, two of six at 8 weeks and one of six at 12 weeks revealed the confirmational change of the H-ras codon 61($A{\rightarrow}T$ transversion). 4. The incidence of point mutation of H-ras codon 12 and 61 were 5.5%(2 of 36) and 11%(4 of 36) respectively. 5. Point mutation of the K-ras could not be seen during the whole experimental period. Form the above results, these findings strongly support the concept that H-ras oncogenes may have the influence of the DMBA induced carcinoma of hamster buccal pouch.

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Photoaging protective effects of BIOGF1K, a compound-K-rich fraction prepared from Panax ginseng

  • Hong, Yo Han;Kim, Donghyun;Nam, Gibaeg;Yoo, Sulgi;Han, Sang Yun;Jeong, Seong-Gu;Kim, Eunji;Jeong, Deok;Yoon, Keejung;Kim, Sunggyu;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.81-89
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    • 2018
  • Background: BIOGF1K, a compound-K-rich fraction, has been shown to display anti-inflammatory activity. Although Panax ginseng is widely used for the prevention of photoaging events induced by UVB irradiation, the effect of BIOGF1K on photoaging has not yet been examined. In this study, we investigated the effects of BIOGF1K on UVB-induced photoaging events. Methods: We analyzed the ability of BIOGF1K to prevent UVB-induced apoptosis, enhance matrix metalloproteinase (MMP) expression, upregulate anti-inflammatory activity, reduce sirtuin 1 expression, and melanin production using reverse transcription-polymerase chain reaction, melanin content assay, tyrosinase assay, and flow cytometry. We also evaluated the effects of BIOGF1K on the activator protein-1 signaling pathway, which plays an important role in photoaging, by immunoblot analysis and luciferase reporter gene assays. Results: Treatment of UVB-irradiated NIH3T3 fibroblasts with BIOGF1K prevented UVB-induced cell death, inhibited apoptosis, suppressed morphological changes, reduced melanin secretion, restored the levels of type I procollagen and sirtuin 1, and prevented mRNA upregulation of MMP-1, MMP-2, and cyclo-oxygenase-2; these effects all occurred in a dose-dependent manner. In addition, BIOGF1K markedly reduced activator-protein-1-mediated luciferase activity and decreased the activity of mitogen-activated protein kinases (extracellular response kinase, p38, and C-Jun N-terminal kinase). Conclusion: Our results strongly suggest that BIOGF1K has anti-photoaging activity and that BIOGF1K could be used in anti-aging cosmeceutical preparations.

Glatiramer acetate inhibits the activation of NFκB in the CNS of experimental autoimmune encephalomyelitis (Glatiramer acetate 투여에 의한 자가면역성 뇌척수염 마우스의 중추신경계에서의 NFκB 활성 억제)

  • Hwang, Insun;Ha, Danbee;Kim, Dae Seung;Joo, Haejin;Jee, Youngheun
    • Korean Journal of Veterinary Research
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    • v.51 no.3
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    • pp.217-225
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    • 2011
  • Glatiramer acetate (GA; Copaxone) has been shown to be effective in preventing and suppressing experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis (MS). It has been recently shown that GA-reactive T cells migrate through the blood-brain barrier, accumulate in the central nervous system (CNS), secrete antiinflammatory cytokines and suppress production of proinflammatory cytokines of EAE and MS. Development of EAE requires coordinated expression of a number of genes involved in the activation and effector functions of inflammatory cells. Activation of inflammatory cells is regulated at the transcriptional level by several families of transcription factors. One of these is the nuclear factor kappa B ($NF{\kappa}B$) family which is present in a variety of cell types and involved in the activation of immune-relative genes during inflammatory process. Since it is highly activated at site of inflammation, $NF{\kappa}B$ activation is also implicated in the pathogenesis of EAE. In this study, we examined whether the inhibition of $NF{\kappa}B$ activation induced by GA can have suppressive therapeutic effects in EAE mice. We observed the expression of $NF{\kappa}B$ and phospho-$I{\kappa}B$ proteins increased in GA-treated EAE mice compared to EAE control groups. The immunoreactivity in inflammatory cells and glial cells of $NF{\kappa}B$ and phospho-$I{\kappa}B$ significantly decreased at the GA-treated EAE mice. These results suggest that treatment of GA in EAE inhibits the activation of $NF{\kappa}B$ and phophorylation of $I{\kappa}B$ in the CNS. Subsequently, the inhibition of $NF{\kappa}B$ activation and $I{\kappa}B$ phosphorylation leads to the anti-inflammatory effects thereby to reduce the progression and severity of EAE.

Effect of Water Extract of Rubi Fructus in RANKL-induced Osteoclast Differentiation (파골세포 분화에 미치는 복분자 물 추출물의 효과)

  • Oh, Jae-Min;Lee, Myeung-Su;Kim, Jeong-Joong;Lee, Jeong-Hugh;Chae, Soo-Uk;Kim, Ha-Young;Jeon, Byung-Hoon;Park, Kie-In;Moon, Seo-Young;Cho, Hae-Joong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.4
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    • pp.669-673
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    • 2011
  • To prevent and treat the osteoporotic fracture, more attention should be paid in old age patients. Osteoclast which has ability to bone resorption is originated from hematopoietic cell line and plays a key role osteoporotic bone loss. Rubi Fructus has been widely used in Oriental medicine. Extracts of the leaves and fruit of Rubus species have been used in various countries as natural remedies to treat diabetes, infections, colic, and burns. However, the effect of extract of Rubi Fructus (fruit of Rubus coreanus Miq.) in osteoclast differentiation remains unknown. Thus, we evaluated the effect of Rubi Fructus on receptor activator of nuclear factor-kB ligand (RANKL)-induced osteoclast differentiation. Here we found that Rubi Fructus significantly inhibited osteoclast differentiation induced by RANKL. Rubi Fructus suppressed the activation of p38 pathway and NFkB in bone marrow macrophages (BMMs) treated with RANKL. Also, Rubi Fructus significantly inhibited the mRNA expression of c-Fos, tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells (NFAT)c1 and cathepsin K in BMMs treated with RANKL. Particularly, Rubi Fructus greatly inhibited the protein expression of c-fos and NFATc1. especially in the case of NFATc1 expression, a master transcription factor of the differentiation of osteoclasts is very important step for osteoclastogenesis. Taken together, our results demonstrated that Rubi Fructus may be useful treatment option of bone-related disease such as osteoporosis and rheumatoid arthritis.

Nuclear Remodeling and In Vitro Development of Bovine Oocytes Following Nuclear Transfer of Bovine Fetal Fibroblasts (태아 섬유아세포로 핵치환된 소 난자의 핵의 재구성과정과 체외 배발달)

  • Um, J. H.;S. J. Uhm;Kim, N-H;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.59-67
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    • 2000
  • This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.

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Physiological Activity of Coffee Beans and Roasted Black Beans (Rhynchosia nulubilis) Mixture Extracts for Coffee Alternative Beverage Development

  • Kim, Ae-Jung;Lee, Hankyu;Ko, Hyuk Wan;Ko, Seong Hee;Woo, Nariyah
    • The Korean Journal of Food And Nutrition
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    • v.29 no.2
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    • pp.178-185
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    • 2016
  • This study was designed to develop and to qualify a coffee alternative beverage using a mixture of coffee beans and roasted black beans (Rhynchosia nulubilis). Therefore, the total isoflavone content (TIC), total phenol content (TPC), antioxidant activity, anti-inflammatory activity, NFATc1 (Nuclear factor of activated T-cells c1) expression in RANKL (receptor activator of nuclear factor kappa-B ligand)-stimulated RAW264.7 cells and sensory evaluation were measured for 5 different Cb (coffee bean)-RoS (roasted seomoktae) mixture extracts (Cb100RoS0, Cb75RoS25, Cb50RoS50, Cb25RoS75, and Cb0RoS100). Cb0RoS100 had the highest TIC ($516.83{\pm}36.61mg/100g$) and TPC ($18.11{\pm}1.77mg$ TAE/100 g) along with the highest antioxidant activity as measured by DPPH radical scavenging activity ($73.55{\pm}8.11%$) and ABTS radical scavenging activity ($63.27{\pm}7.27%$). Also, Cb0RoS100 showed the highest anti-inflammatory activity as measured by NO production ($13.57{\pm}2.21{\mu}M$) and PGE2 production ($3.25{\pm}0.21ng/mL$). The more the RoS ratio was increased in the mixtures of Cb-RoS, the more the NFATc1 protein expression was decreased in RANKL-stimulated RAW264.7 cells. In case of sensory evaluation, Cb50RoS50 had the highest scores for flavor, delicate flavor and overall quality, which were similar to those in Cb alone (Cb100RoS0). We suggest that the use of RoS replacement instead of Cb in/as a coffee alternative beverage may help to reduce the risk of caffeine-related bone loss and/or bone disease by effectively blocking NFATc1 expression in RANKL-stimulated RAW264.7 cells compared with Cb alone.

Nuclear Transfer using Human CD59 and IL-18BP Double Transgenic Fetal Fibroblasts in Miniature Pigs

  • Ryu, Junghyun;Kim, Minjeong;Ahn, Jin Seop;Ahn, Kwang Sung;Shim, Hosup
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.1-7
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    • 2016
  • Xenotransplantation involves multiple steps of immune rejection. The present study was designed to produce nuclear transfer embryos, prior to the production of transgenic pigs, using fibroblasts carrying transgenes human complement regulatory protein hCD59 and interleukin-18 binding protein (hIL-18BP) to reduce hyperacute rejection (HAR) and cellular rejection in pig-to-human xenotransplantation. In addition to the hCD59-mediated reduction of HAR, hIL-18BP may prevent cellular rejection by inhibiting the activation of natural killer cells, activated T-cell proliferation, and induction of $IFN-{\gamma}$. Transgene construct including hCD59 and ILI-18BP was introduced into miniature pig fetal fibroblasts. After antibiotic selection of double transgenic fibroblasts, integration of the transgene was screened by PCR, and the transgene expression was confirmed by RT-PCR. Treatment of human serum did not affect the survival of double-transgenic fibroblasts, whereas the treatment significantly reduced the survival of non-transgenic fibroblasts (p<0.01), suggesting alleviation of HAR. Among 337 reconstituted oocytes produced by nuclear transfer using the double transgenic fibroblasts, 28 (15.3%) developed to the blastocyst stage. Analysis of individual embryos indicated that 53.6% (15/28) of embryos contained the transgene. The result of the present study demonstrates the resistance of hCD59 and IL-18BP double-transgenic fibroblasts against HAR, and the usefulness of the transgenic approach may be predicted by RT-PCR and cytolytic assessment prior to actual production of transgenic pigs. Further study on the transfer of these embryos to surrogates may produce transgenic clone miniature pigs expressing hCD59 and hIL-18BP for xenotransplantation.

Sex Determination in Somatic and Embryonic Cells of the Pig by FISH and PCR (FISH와 PCR에 의한 돼지 체세포 및 배아세포의 성 판정)

  • Chung, Y.;Jeon, J.T.;Kim, K.D.;Lee, S.H.;Hong, K.C.
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.323-331
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    • 1996
  • Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

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